Autoreactive CD8+ T cell exhaustion distinguishes subjects with slow type 1 diabetes progression
Alice E. Wiedeman, Virginia S. Muir, Mario G. Rosasco, Hannah A. DeBerg, Scott Presnell, Bertrand Haas, Matthew J. Dufort, Cate Speake, Carla J. Greenbaum, Elisavet Serti, Gerald T. Nepom, Gabriele Blahnik, Anna M. Kus, Eddie A. James, Peter S. Linsley, S. Alice Long
Alice E. Wiedeman, Virginia S. Muir, Mario G. Rosasco, Hannah A. DeBerg, Scott Presnell, Bertrand Haas, Matthew J. Dufort, Cate Speake, Carla J. Greenbaum, Elisavet Serti, Gerald T. Nepom, Gabriele Blahnik, Anna M. Kus, Eddie A. James, Peter S. Linsley, S. Alice Long
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Research Article Autoimmunity Immunology

Autoreactive CD8+ T cell exhaustion distinguishes subjects with slow type 1 diabetes progression

Abstract

Although most patients with type 1 diabetes (T1D) retain some functional insulin-producing islet β cells at the time of diagnosis, the rate of further β cell loss varies across individuals. It is not clear what drives this differential progression rate. CD8+ T cells have been implicated in the autoimmune destruction of β cells. Here, we addressed whether the phenotype and function of autoreactive CD8+ T cells influence disease progression. We identified islet-specific CD8+ T cells using high-content, single-cell mass cytometry in combination with peptide-loaded MHC tetramer staining. We applied a new analytical method, DISCOV-R, to characterize these rare subsets. Autoreactive T cells were phenotypically heterogeneous, and their phenotype differed by rate of disease progression. Activated islet-specific CD8+ memory T cells were prevalent in subjects with T1D who experienced rapid loss of C-peptide; in contrast, slow disease progression was associated with an exhaustion-like profile, with expression of multiple inhibitory receptors, limited cytokine production, and reduced proliferative capacity. This relationship between properties of autoreactive CD8+ T cells and the rate of T1D disease progression after onset make these phenotypes attractive putative biomarkers of disease trajectory and treatment response and reveal potential targets for therapeutic intervention.

Authors

Alice E. Wiedeman, Virginia S. Muir, Mario G. Rosasco, Hannah A. DeBerg, Scott Presnell, Bertrand Haas, Matthew J. Dufort, Cate Speake, Carla J. Greenbaum, Elisavet Serti, Gerald T. Nepom, Gabriele Blahnik, Anna M. Kus, Eddie A. James, Peter S. Linsley, S. Alice Long

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Figure 5

Islet-specific CD8+ T cells with an abundant cluster 1 (exhausted) phenotype are hypoproliferative and produce limited levels of the cytokines IL-2 and IFN-γ.

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Islet-specific CD8+ T cells with an abundant cluster 1 (exhausted) pheno...
PBMCs from individuals with T1D (n = 11) with varying frequencies of cluster 1 among their islet-specific cells were stimulated with anti-CD3 plus anti-CD28. Cells were assayed by flow cytometry to identify islet-specific (Tmr+) CD8+ T cells (Supplemental Figure 13). Examples of gating for proliferation and cytokine production are shown for a rapid progressor (T1D-02) and a slow progressor (T1D-34) with low (4%) and high (60%) frequencies of cluster 1, respectively. (A) Representative examples of the frequency of proliferated cells on day 5 among stimulated (black line) islet Tmr+ cells as measured by CellTrace dye dilution, using unstimulated (solid gray) cells as a gating control. (B) Frequency of proliferated cells among islet Tmr+ cells after 5 days of stimulation, plotted against the frequency of cluster 1 determined by mass cytometry for each individual (n = 11). (C) Representative examples of IL-2 and IFN-γ production assessed at 6 hours among islet Tmr+ (black) or Tmr– CD8+ T cells (gray). (D) Frequency of IL-2+ and IFN-γ+ cells among islet Tmr+ cells after 6 hours of stimulation, plotted against the frequency of cluster 1 determined by mass cytometry for each individual (n = 10); no substantial cytokine production (<1%) was observed in the absence of stimulation. Statistical significance was determined by Spearman’s correlation. The difference in proliferation between islet-specific cells of rapid progressors (triangles, n = 3) and slow progressors (squares, n = 4) was not significant (P = 0.057), nor was cytokine production (P > 0.05) by Mann-Whitney U test.